This study examined plasma proline concentration flux, oxidation, and endogenous biosynthesis in five healthy young men given three isocaloric, isonitrogenous diets for 1 wk [a complete egg-pattern amino acid diet (diet 1), an amino acid mixture devoid of proline (diet 2), and a diet composed solely of indispensable amino acids (diet 3)]. At the end of each dietary period, a 360-min postabsorptive, primed, continuous stable-isotope-tracer infusion of L-[1-13C]proline and L-[methyl-2H3]leucine was performed in all subjects. Plasma proline concentrations declined by 22% on diet 2 (p less than 0.02) and by 29% on diet 3 (p less than 0.01). No statistically significant (p greater than 0.2) changes were observed for proline oxidation, endogenous biosynthesis, or flux. The data suggest that the absence of proline in the human diet does not trigger changes in proline dynamics during the postabsorptive state. The metabolic significance of the reduction of plasma proline concentrations requires elucidation.